Evaluation of cell surface antigens as potential targets for recombinant tumor toxins.
Tumor toxins are recombinant polypeptides derived from the fusion of a single-chain antibody domain or a ligand specific for a cell surface receptor with the enzymatic domain of a bacterial or plant toxin. If these receptors are preferentially expressed on the surface of tumor cells, the tumor toxins can serve as efficient therapeutics. Examples in which growth factors, cytokines, hormones or antibody domains have been fused to toxins have been described and the bacterially expressed molecules have shown anti-tumor effects in vitro and in vivo. Since many tumors are not sensitive to the existing tumor toxins, the search continues for new antigens with tumor specific or tumor enhanced expression. We explored three antigenic structures for their possible use as targets for tumor toxins, i.e. (a) a glycosphingolipid with an Le(a) antigenic structure which is overexpressed in gastrointestinal carcinomas, e.g. colon carcinoma, (b) the epithelial glycoprotein episialin which shows altered expression in various tumor tissues such as breast, ovary and pancreas and (c) the neurotensin receptor which is found on NSLC, pancreatic and colonic cancer cells. For targeting the tumor toxin to the carbohydrate structures, we constructed recombinant single-chain antibody domains (scFv) fused to a truncated form of the exotoxin A from Pseudomonas aeruginosa (ETA). In the case of the neurotensin receptor, the ligand neurotensin was exploited to construct a recombinant fusion protein with exotoxin A as the effector domain. Binding experiments showed that the recombinant proteins are able to specifically recognize their respective target structures. In vitro cytotoxicity assays with neurotensin-ETA showed specific killing activity for receptor positive tumor cell lines. For the scFv-ETA constructs we could not observe in vitro tumor toxicity. We conclude that the antigenic targets for tumor toxins have to be carefully chosen and that specific cell binding activity is not a sufficient criterion for tumor cell killing. The neurotensin-ETA, a new anti-tumor agent, shows that seven transmembrane spanning receptors are potential targets which allow toxin binding and promote cell killing through the appropriate internalization.
Wick
B
B
Institute for Experimental Cancer Research, Tumor Biology Center, Freiburg, Germany.
Groner
B
B
D036002:ADP Ribose Transferases; D000818:Animals; D000954:Antigens, Surface; D001427:Bacterial Toxins; D005098:Exotoxins / Q000008:administration & dosage*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000276:immunology; D000922:Immunotoxins / Q000008:administration & dosage*; D008024:Ligands; D051379:Mice; D018396:Mucin-1 / Q000276:immunology; D009496:Neurotensin / Q000276:immunology; D011485:Protein Binding; D018028:Receptors, Neurotensin / Q000276:immunology; D011994:Recombinant Proteins; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D000954:Antigens, Surface; D001427:Bacterial Toxins; D005098:Exotoxins; D007128:Immunoglobulin Fragments; D000922:Immunotoxins; D008024:Ligands; D018396:Mucin-1; D009496:Neurotensin; D018028:Receptors, Neurotensin; D011994:Recombinant Proteins; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A